Stable compositions of peptide epoxy ketones

ABSTRACT

The invention relates to pharmaceutical compositions that provide improved solubility and stability for peptide epoxy ketones. More specifically, the invention relates to pharmaceutical compositions comprising the peptide epoxy ketone proteasome inhibitor carfilzomib.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/699,752, filed Sep. 11, 2012, the specification of which is herebyincorporated by reference in its entirety.

FIELD OF INVENTION

The present invention relates to pharmaceutical compositions comprisinga peptide epoxy ketone, which can be a proteasome inhibitor, such asCarfilzomib, wherein the pharmaceutical compositions provide improvedstability and solubility for the proteasome inhibitor contained therein.

BACKGROUND OF THE INVENTION

Carfilzomib, which is the active ingredient in KYPROLIS®, is a peptideepoxy ketone proteasome inhibitor indicated for the treatment ofpatients with multiple myeloma who have received at least two priortherapies, including bortezomib and an immunomodulatory agent, and havedemonstrated disease progression on or within 60 days of completion ofthe last therapy. Carfilzomib irreversibly binds to the N-terminalthreonine-containing active sites of the 20S proteasome, the proteolyticcore particle within the 26S proteasome. Carfilzomib has been shown tohave antiproliferative and proapoptotic activities in vitro in solid andhematologic tumor cells. The compound has also been shown to inhibitproteasome activity in blood and tissue and delay tumor growth in modelsof multiple myeloma, hematologic, and solid tumors.

Peptide epoxy ketone proteasome inhibitors, such as Carfilzomib, haveproven difficult to formulate due to their low aqueous solubility. Inaddition, the proteasome inhibitors are very unstable. Thus,compositions comprising these compounds must typically be lyophilizedbefore storage and reconstituted before use. Moreover, the reconstitutedcompositions are often not stable themselves (even under refrigeratedconditions). As shown in Table 1, certain reconstituted KYPROLIS®compositions must be used within 24 hours after reconstitution becauseof their instability.

TABLE 1 Storage Conditions of Reconstituted Stability*per ContainerKYPROLIS Vial Syringe IV Bag (¶) Refrigerated (2° C. to 8° C.; 24 hours24 hours 24 hours 36° F. to 46° F.) Room Temperature (15° C. to 30° C.; 4 hours  4 hours  4 hours 59° F. to 86° F.) *Total time fromreconstitution to administration should not exceed 24 hours. ¶ 5%Dextrose Injection, USP

The need to lyophilize a composition comprising a peptide epoxy ketoneproteasome inhibitor presents challenges to healthcare professionals.For example, dosing errors frequently occur during reconstitution ofsuch products, and safety risks are presented if the reconstitution isnot conducted aseptically. Additionally, the product afterreconstitution is not stable for more than 24 hr, a product with suchstability and storage temperature restrictions poses a problem forconvenient use in a clinical use setting. Furthermore, lyophilizedproducts allow for a one time reconstitution and use per patient. Thistype of presentation does not allow for injecting multiple doses. Alsoany unused portion of the reconstituted composition must be discarded,which leads to drug wasting. For at least these reasons, there is a needfor new pharmaceutical compositions of peptide epoxy ketone proteasomeinhibitors that provide improved solubility and stability for thepeptide epoxy ketone proteasome inhibitor, as well as for ready-to-usecompositions that eliminate the need for reconstitution.

SUMMARY OF THE INVENTION

Accordingly, in one aspect, the invention provides a lyophilizedcomposition comprising a peptide epoxy ketone and a non-volatile sugaracid.

In one embodiment, the non-volatile sugar acid is lactobionic acid.

In one embodiment, the peptide epoxy ketone is a proteasome inhibitor.

In one embodiment, the peptide epoxy ketone is Carfilzomib.

In one embodiment, the peptide epoxy ketone is Carfilzomib and thenon-volatile sugar acid is lactobionic acid.

The lyophilized pharmaceutical composition may be administered to apatient in need of such compositions after the composition isreconstituted with one or more pharmaceutically acceptable diluents.

In another aspect, the invention provides a pharmaceutical compositioncomprising (i) a peptide epoxy ketone; (ii) a solvent system comprisingone or more water miscible pharmaceutically acceptable organic solventssuitable for injection, and optionally water; and (iii) a non-volatilesugar acid.

In one embodiment, the water miscible pharmaceutically acceptableorganic solvent suitable for injection is selected from the groupconsisting of ethanol, propylene glycol, polyethylene glycol, glycerol,dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, and mixturesthereof.

In one embodiment, the water miscible pharmaceutically acceptableorganic solvent suitable for injection is selected from the groupconsisting of ethanol, propylene glycol, polyethylene glycol, andmixtures thereof

In one embodiment, the non-volatile sugar acid is lactobionic acid.

In one embodiment, the peptide epoxy ketone is a proteasome inhibitor.

In one embodiment, the peptide epoxy ketone is Carfilzomib.

In one embodiment, the peptide epoxy ketone is Carfilzomib and thenon-volatile sugar acid is lactobionic acid.

In one embodiment, the peptide epoxy ketone is Carfilzomib; thenon-volatile sugar acid is lactobionic acid; and the water misciblepharmaceutically acceptable organic solvent suitable for injection isselected from the group consisting of ethanol, propylene glycol,polyethylene glycol, and mixtures thereof

In one embodiment, the pharmaceutical composition is a ready to useformulation.

In one embodiment, the pharmaceutical composition is a ready to diluteformulation.

The pharmaceutical compositions of the invention may also comprise oneor more pharmaceutically acceptable excipients, such as a buffer,surfactant, antioxidant, preservative, isotonicity agent, and/or alyoprotectant agent.

Another aspect of the invention relates to methods of making thepharmaceutical compositions discussed herein.

The pharmaceutical compositions of the invention overcome one or morelimitations discussed above.

DETAILED DESCRIPTION OF THE INVENTION

The invention is generally directed towards pharmaceutical compositionscomprising a peptide epoxy ketone.

In one embodiment, the composition is a lyophilized compositioncomprising the peptide epoxy ketone and a non-volatile sugar acid.

In one embodiment, the composition is a liquid formulation comprising(i) a peptide epoxy ketone; (ii) a solvent system comprising one or morewater miscible pharmaceutically acceptable organic solvents suitable forinjection, and optionally water; and (iii) a non-volatile sugar acid.

In each of the embodiments, the peptide epoxy ketone can be a proteasomeinhibitor.

In each of the embodiments, the peptide epoxy ketone can be Carfilzomib.

Where such compositions are lyophilized or concentrated above theconcentration suitable for injection or infusion, the compositions willbe administered after reconstitution with one or more pharmaceuticallyacceptable diluents.

Regardless of the particular form of the preparation, the pharmaceuticalcompositions contemplated herein may further comprise one or morebuffers, surfactants, antioxidants, preservatives, isotonicity agents,and/or lyoprotectants.

Pharmaceutical compositions according to the instant disclosure provideimproved stability and solubility for peptide epoxy ketones, such asCarfilzomib.

Any recitation of ranges of values set forth below is merely intended toserve as a shorthand method of referring individually to each separatevalue falling within the range, unless otherwise indicated herein, andeach separate value is incorporated into the specification as if it wereindividually recited herein. Furthermore, all references, includingpublications, patent applications, and patents, cited herein are herebyincorporated by reference to the same extent as if each reference wereindividually and specifically indicated to be incorporated by referenceand were set forth in its entirety herein.

The terms “a” and “an” and “the,” as used herein, are to be construed tocover both the singular and the plural, unless otherwise indicatedherein or clearly contradicted by context.

The terms “comprising,” “having,” “including,” and “containing,” as usedherein, are to be construed as open-ended terms (i.e., meaning“including, but not limited to,”) unless otherwise noted.

The pharmaceutical compositions of the invention comprise at least onepeptide epoxy ketone or a pharmaceutically acceptable salt thereof.Preferably, the peptide epoxy ketone is a proteasome inhibitor. Peptideepoxy ketones are epoxide-containing compounds, which contain a ketonegroup proximate to an epoxide group. Peptide epoxy ketones have thegeneral structure:

wherein:

R¹ and R² are hydrogen or a C₁-C₈ alkyl group, such as methyl, ethyl,propyl and butyl groups, which can be further substituted with one ormore hydroxy, halogen, amino, carboxy, carbonyl, thio, sulfide, ester,amide, or C₁-C₆ alkoxy groups; and

P is a peptide chain containing between 1 and 12 amino acids that isconnected to the nitrogen by an amide bond with the carboxylic acidterminus of the amino acid. The N-terminal of the peptide chain nay beprotected or derivatized.

The stereochemistry of the carbons bonded to R¹ and R² can be (R) or(S). In one preferred embodiment the stereochemistry of the carbonbonded to R¹ and R² is as depicted below:

For some embodiments, the oxygen atom of the epoxide is configured asdepicted above in order to facilitate interaction with an N-terminalnucleophilic group in an Ntn hydrolase. For example, irreversibleinteractions of enzyme inhibitors with the β5/Pre2 subunit of 20Sproteasome, which lead to inhibition, appear to be facilitated by theconfiguration illustrated above. In the case of other Ntn hydrolases,the opposite stereochemistry of the epoxide may be preferred.

Preferably, the peptide chain contains two or more amino acids. In someembodiments, useful for inhibiting chymotrypsin-like (CT-L) activity ofthe proteasome, the peptide chain contains between two and eight aminoacids, and in some preferred embodiments for CT-L inhibition, betweentwo and six amino acids. In other embodiments useful for inhibiting thePGPH activity of the proteasome, the peptide chain contains between twoand eight amino acids, and in some preferred embodiments for PGPHinhibition, between two and six amino acids. In other embodiments, thepeptide chain contains between two and four amino acid units.

The amino acids can be a natural occurring essential amino acid, such asglycine, alanine, valine, isoleucine, leucine, phenylalanine, threonine,tyrosine, serine, and proline. The amino acids can be a naturallyoccurring non-essential amino acid, such as for example taurine,carnitine, citrulline, cystine, omithine, norleucine, and others. Theamino acids can also be a non-naturally occurring amino acid of generalformula:

wherein the side chain, R, can be hydrogen, phenyl, or C₁-C₈ alkyl,wherein the C₁-C₈ alkyl can be further substituted with one or morephenyl, hydroxy, halogen, amino, carboxy, carbonyl, thio, sulfide,ester, amide, or C₁-C₆ alkoxy groups, and the phenyl can be furthersubstituted with one or more hydroxy, halogen, amino, carboxy, carbonyl,thio, sulfide, ester, amide, or C₁-C₆ alkoxy groups.

Peptide epoxy ketones useful in the compositions of the inventioninclude those described in U.S. Pat. No. 7,737,112, the content of whichare incorporated herein by reference.

Most preferably, the peptide epoxy ketone proteasome inhibitor isCarfilzomib (i.e.,(2S)-N-((S)-1-(S)-4-methyl-1-((R)-2-methyloxiran-2-yl)-1-oxopentan-2-ylcarbamoyl)-2-phenylethyl)-2-((S)-2-(2-morpholinoacetamido)-4-phenylbutanamido)-4-methylpentanamide)which has the following structure:

Lyophilized Compositions:

The lyophilized compositions of the invention comprise the peptide epoxyketone and a non-volatile sugar acid. The peptide epoxy ketone may be aproteasome inhibitor. Preferably, the peptide epoxy ketone isCarfilzomib.

The weight ratio of the non-volatile sugar acid to the peptide epoxyketone ranges from about 2:1 to 15:1, preferably about 3:1 to 15:1, morepreferably about 4:1 to 15:1, most preferably about 4:1 to 10:1.Typically, the weight ratio of the sugar acid to the peptide epoxyketone is greater than 4:1, preferably greater than 5:1. Typically, theweight ratio of the sugar acid to the peptide epoxy ketone is less than15:1, preferably less than 10:1.

In one embodiment, the lyophilized composition comprises about 60 mg ofthe peptide epoxy ketone and about 300 mg of the non-volatile sugaracid. In one embodiment, the lyophilized composition comprises about 60mg of Carfilzomib and about 300 mg of lactobionic acid. In oneembodiment, the lyophilized composition comprises about 60 mg ofCarfilzomib and about 400 mg of lactobionic acid.

The phrase sugar acid means an oxidized derivative of sugar having oneor more carboxylic acid functional groups. Sugar acids can be obtainedby oxidizing the aldehyde group of a sugar. The term sugar includes anypolyhydroxy carbohydrate moiety, including monosaccharides,disaccharides, polysaccharides. Preferably, the sugar acid is derivedfrom a monosaccharide or a disaccharide. In one embodiment the sugaracid is a monosaccharide. In one embodiment the sugar acid is adisaccharide. The sugar acids may be used in the acid form or as a salt.When used as a salt, the salt is preferably a sodium, calcium,magnesium, or zinc salt. The sugar acid may be in the form of an esteror a lactone. Sugar acids also include compounds derived from aminosugars and acetylated amino sugars.

Non-volatile sugar acids useful in the compositions of the inventioninclude, but are not limited to, N-acetylneuraminic acid,N-acetyltalosaminuronic acid, aldaric acid, aldonic acid,3-deoxy-D-manno-oct-2-ulosonic acid, galaturonic acid, D-galacturonicacid, glucaric acid, gluconic acid, glucuronic acid, glucono-γ-lactone,glyceric acid, N-glycolylneuraminic acid, iduronic acid, isosaccharinicacid, lactobionic acid, mucic acid, muramic acid, neuraminic acid,pangamic acid, saccharic acid, sialic acid, threonic acid, ulosonicacid, uronic acid, X-Gluc, xylonic acid, ascorbic acid, and mixturesthereof. Preferably, the non-volatile organic acid is lactobionic acid.

Preferred sugar acids include, but are not limited to, gluconic acid,glucono-γ-lactone, lactobionic acid, glucuronic acid and its mono-ordilactones, pangamic acid, mannosaccharic acid and its mono-ordilactones, mucic acid and its mono-or dilactones, and mixtures thereof.Most preferred sugar acid is lactobionic acid.

The lyophilized compositions may further include one or more excipientssuitable for inclusion in a lyophilized composition. For example, thelyophilized compositions may include a lyoprotectant. Suitablelyoprotectants include amino acids and polymers. Preferably, the aminoacid is selected from the group consisting of lysine, alanine, andglycine. Suitable polymers include various proteins (e.g., gelatin andalbumin), polyethylene glycol, gelatin, polyvinyl pyrrolidone, albumin,and Dextran-40. Typically the lyoprotectant represents less than 50%weight/weight of the lyophilized composition, and all concentrationsabove 1% weight/weight of the total composition are deemed effective toenhance the stability of the composition. In various embodiments, thelyoprotectant is present in an amount of at least about 5%weight/weight, at least about 10% weight/weight, or at least about 20%weight/weight of the total composition.

The compositions can be prepared by combining the peptide epoxy ketone,the non-volatile sugar acid, any optional excipients, and a suitablelyophilization solvent to provide a solution and then lyophilizing thesolution using art-recognized lyophilization techniques.

The compositions can be sterilized. Sterilized compositions can beobtained using art-recognized methods for sterilization, such asfiltration through 0.22 micron filters (e.g., PVDF filters), heatsterilization, radiation (e.g., gamma, electron beam, microwave), and/orethylene oxide sterilization.

The non-volatile sugar acid compositions are more stable than acomparable composition that does not include the non-volatile sugaracid. Preferably the compositions have sufficient stability to allowstorage at a commercially relevant temperature, such as between about 0°C. and about 60° C., for a commercially relevant period of time, such asat least one week, preferably at least one month, more preferably atleast three months, and most preferably at least six months. In someembodiments the compositions have sufficient stability to allow storageat a commercially relevant temperature for more than one year, and insome embodiments more than two years. Stability can be measured usingany physiochemical characterization techniques known to those skilled inthe art, such as, for example high pressure liquid chromatography(HPLC).

The lyophilized pharmaceutical compositions discussed herein may be in alyophilized powder or lyophilized cake form. Before use, the lyophilizedpharmaceutical compositions are diluted or reconstituted with apharmaceutically acceptable diluent. Any diluent known in the art inwhich the peptide epoxy ketone is soluble can be used. Preferably, thelyophilized composition is reconstituted with a solvent system so as toprovide a liquid pharmaceutical composition as described below. In oneembodiment, reconstitution provides a ready to use formulation asdescribed below. In one embodiment, reconstitution provides a ready todilute formulation as described below. The ready to dilute formulationcan then be diluted with a suitable diluent before administration.Suitable diluents include, but are not limited to, water, saline,dextrose 5% in water, water for injection, and lactated ringer'ssolution. In one embodiment, the lyophilized composition isreconstituted directly with a suitable diluent.

The lyophilized pharmaceutical compositions are preferably contained ina vial or pre-filled syringe before reconstitution occurs.

It should be appreciated that the peptide epoxy ketone is present in thelyophilized pharmaceutical compositions in an amount that is suitablefor administration after reconstitution.

Liquid Pharmaceutical Compositions:

The liquid pharmaceutical compositions of the invention comprise (i) apeptide epoxy ketone, (ii) a solvent system comprising one or more watermiscible pharmaceutically acceptable organic solvents suitable forinjection and optionally water, and (iii) a non-volatile sugar acid.

In one embodiment, the liquid pharmaceutical composition is a ready touse formulation. A ready to use formulation is a formulation that issuitable for administration without further dilution.

In one embodiment, the liquid pharmaceutical composition is a ready todilute formulation. A ready to dilute formulation is a formulation thatis suitable for administration after dilution with a suitable diluent.

The liquid pharmaceutical compositions can be a multi-dose formulation,i.e., a formulation that has a volume and/or quantity of the activepharmaceutical ingredient suitable for at least two independent anddistinct administrations (to the same or a different patient) of theformulation.

The liquid pharmaceutical compositions are more stable than a comparablecomposition that does not include the non-volatile sugar acid. Withoutwishing to be bound by theory, it is believed that the sugar acidprovides improved stability to the liquid pharmaceutical compositions.Importantly, the sugar acid also helps prevent the peptide epoxy ketonefrom precipitating when the liquid pharmaceutical composition is furtherdiluted with a diluent. Thus, the liquid pharmaceutical compositions canadvantageously be more readily diluted with a suitable diluent, withoutthe peptide epoxy ketone precipitating. By reducing the tendency of thepeptide epoxy ketone to precipitate, permits the preparation of moreconcentrated formulations to be obtained by diluting the by liquidpharmaceutical compositions.

The water miscible pharmaceutically acceptable organic solvent suitablefor injection provides improved solubility for the peptide epoxy ketonecompared to an aqueous composition in the absence of thepharmaceutically acceptable organic solvent. The pharmaceuticallyacceptable organic solvent also provides a solvent system that reducesdegradation of the peptide epoxy ketone compared to an aqueous solventin the absence of the organic solvent

In one embodiment, the peptide epoxy ketone is a proteasome inhibitor.In one embodiment, the peptide epoxy ketone is Carfilzomib.

Typically, the concentration of the peptide epoxy ketone in the liquidpharmaceutical composition ranges from about 0.1 to about 10.0 mg/mL.For example, the concentration of the peptide epoxy ketone in a ready touse formulation ranges typically from about 0.1 to about 3.0 mg/mL andthe concentration of the peptide epoxy ketone in a ready to diluteformulation typically ranges from about 1.0 to about 10.0 mg/mL.

Any of the non-volatile sugar acids described above can be used in theliquid pharmaceutical compositions. The weight ratio of the non-volatilesugar acid to the peptide epoxy ketone ranges from about 2:1 to 15:1,preferably about 3:1 to 15:1, more preferably about 4:1 to 15:1, mostpreferably about 4:1 to 10:1. Typically, the weight ratio of thenon-volatile sugar acid to the peptide epoxy ketone is greater than 4:1,preferably greater than 5:1. Typically, the weight ratio of thenon-volatile sugar acid to the peptide epoxy ketone is less than 15:1,preferably less than 10:1.

Preferably, the non-volatile sugar acid is lactobionic acid.

Suitable water miscible pharmaceutically acceptable organic solventssuitable for injection are non-toxic organic solvents in which thepeptide epoxy ketone is soluble. Suitable solvents include of ethanol,propylene glycol, polyethylene glycol, glycerol, dimethylacetamide,N-methylpyrrolidone, dimethylsulfoxide, and mixtures thereof In oneembodiment, the solvent is selected from the group consisting ofethanol, propylene glycol, polyethylene glycol, and mixtures thereof.

Preferably, the water miscible pharmaceutically acceptable organicsolvent suitable for injection is selected from the group consisting ofethanol, propylene glycol, polyethylene glycol (in particular PEG 300and PEG 400), and mixtures thereof

The liquid pharmaceutical compositions may include water. Preferably thewater miscible pharmaceutically acceptable organic solvent suitable forinjection comprises more than about 25% by volume of the total amount ofliquid used to formulate the pharmaceutical composition. Morepreferably, the water miscible pharmaceutically acceptable organicsolvent suitable for injection comprises more than about 50% by volumeof the total amount of liquid used to formulate the pharmaceuticalcomposition. Most preferably, the water miscible pharmaceuticallyacceptable organic solvent suitable for injection comprises more thanabout 75% by volume of the total amount of liquid used to formulate thepharmaceutical compositions

In one embodiment, the liquid pharmaceutical composition comprises atleast two pharmaceutically acceptable solvents. The at least twopharmaceutically acceptable solvents can be present in any ratio.

In one embodiment, the liquid pharmaceutical composition comprises bothethanol and propylene glycol. Preferably, the volume ratio of ethanol topropylene glycol ranges from about 1:10 to 10:1. In one embodiment, theliquid pharmaceutical composition comprises ethanol, propylene glycol,and water. In one embodiment, the volume percent of the water rangesfrom 0% to about 60% of the total volume of liquid in the composition.

In one embodiment, the liquid pharmaceutical composition comprises bothethanol and polyethylene glycol (e.g., PEG 300 or PEG 400). Preferably,the volume ratio of ethanol to polyethylene glycol ranges from about1:10 to 10:1. In one embodiment, the liquid pharmaceutical compositioncomprises ethanol, polyethylene glycol, and water. In one embodiment,the volume percent of the water ranges from 0% to about 60% of the totalvolume of liquid in the composition.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib; a solvent system comprising ethanol, propylene glycol, andwater; and a non-volatile sugar acid.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib; a solvent system comprising ethanol, propylene glycol, andwater; and lactobionic acid.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, propylene glycol, and water; and lactobionic acid.In one embodiment, the Carfilzomib is present in an amount of about 1 to5 mg/mL.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, propylene glycol, and water; and lactobionic acid,wherein the weight ratio of lactobionic acid to Carfilzomib ranges fromabout 2:1 to about 15:1. In one embodiment, the Carfilzomib is presentin an amount of about 1 to 5 mg/mL.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, propylene glycol, and water; and lactobionic acid,wherein the weight ratio of lactobionic acid to Carfilzomib ranges fromabout 2:1 to about 15:1, and the ratio of propylene glycol to ethanol isabout 1:10 to 10:1 (v/v). In one embodiment, the ethanol and propyleneglycol comprise more than about 25% by volume of the total amount ofliquid used to formulate the pharmaceutical composition. In oneembodiment, the ethanol and propylene glycol comprise more than about50% by volume of the total amount of liquid used to formulate thepharmaceutical composition. In one embodiment, the ethanol and propyleneglycol comprise more than about 75% by volume of the total amount ofliquid used to formulate the pharmaceutical composition. In oneembodiment, the Carfilzomib is present in an amount of about 1 to 5mg/mL.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib; a solvent system comprising ethanol, polyethylene glycol,and water; and a non-volatile sugar acid. In one embodiment, thepolyethylene glycol is selected from PEG 300 and PEG 400.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib; a solvent system comprising ethanol, polyethylene glycol,and water; and lactobionic acid. In one embodiment, the polyethyleneglycol is selected from PEG 300 and PEG 400.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, polyethylene glycol, and water; and lactobionicacid. In one embodiment, the polyethylene glycol is selected from PEG300 and PEG 400. In one embodiment, the Carfilzomib is present in anamount of about 1 to 5 mg/mL.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, polyethylene glycol, and water; and lactobionicacid, wherein the weight ratio of lactobionic acid to Carfilzomib rangesfrom about 2:1 to about 15:1. In one embodiment, the polyethylene glycolis selected from PEG 300 and PEG 400.

In one embodiment, the liquid pharmaceutical composition comprisesCarfilzomib in an amount of about 0.1 to 10 mg/mL; a solvent systemcomprising ethanol, polyethylene glycol, and water; and lactobionicacid, wherein the weight ratio of Carfilzomib to lactobionic acid rangesfrom about 2:1 to about 15:1, and the ratio of ethanol to polyethyleneglycol is about 1:10 to 10:1 (v/v). In one embodiment, the ethanol andpolyethylene glycol comprise more than about 25% by volume of the totalamount of liquid used to formulate the pharmaceutical composition. Inone embodiment, the ethanol and polyethylene glycol comprise more thanabout 50% by volume of the total amount of liquid used to formulate thepharmaceutical composition. In one embodiment, the ethanol andpolyethylene glycol comprise more than about 75% by volume of the totalamount of liquid used to formulate the pharmaceutical composition. Inone embodiment, the Carfilzomib is present in an amount of about 1 to 5mg/mL.

The liquid pharmaceutical compositions of the instant invention mayfurther comprise one or more excipients, such as, buffers, surfactants,antioxidants, preservatives, isotonicity agents, and/or lyoprotectants.

For instance, a buffer is typically used in the liquid pharmaceuticalcompositions of the invention to control the pH. As shown in Table 2, atleast some peptide epoxy ketones (e.g., Carfilzomib) have demonstrated astrong pH-dependent solubility.

TABLE 2 Effect of pH on Solubility of Carfilzomib pH SaturationSolubility (μg/mL) 1.7 (1% L-Cysteine) 459.0 3.5 (15 mM Acetate Buffer)125.0 5.0 (15 mM Citrate Buffer) 9.0 7.0 (15 mM Phosphate Buffer) 5.0Accordingly, a buffer may be used in the liquid pharmaceuticalcompositions of the instant invention to maintain a pre-selected pHlevel at which the peptide epoxy ketone is soluble.

Examples of suitable buffers include mixtures of a weak acid and alkalimetal salt (e.g., sodium, potassium) and the conjugate base of the weakacid. Suitable buffers include, for example, buffers selected from thegroup consisting of citric acid, acetic acid, maleic acid, phosphoricacid, succinic acid, or tartaric acid, as well as the counter ion saltsthereof. The molar concentration of the buffer typically ranges betweenabout 5 millimolar and about 150 millimolar, and is most preferablyabout 15 millimolar. The buffer will typically have a pH of about 3.5,however, modifications to the pH value are also contemplated herein. Itwill be recognized by those of skill in the art that determination ofthe preferred solubility range of a peptide epoxy ketone around a pH of3.5 using any of the aforementioned buffers can be performed using knowntechniques.

The pH level for each pharmaceutical composition should be selected toprovide suitable solubility of the peptide epoxy ketone used therein. Itis generally preferred, however, that the pH of the compositions besuitable for injection and, therefore, will typically be between about2.0 and about 9.0, and even more typically between about 2.5 and about8.0.

The liquid pharmaceutical compositions according the invention may alsocomprise an anionic, nonionic, cationic, or amphoteric surfactant.Anionic surfactants that may be used in the pharmaceutical compositionsof the invention include alkyl ether sulfates and carboxylic acids, inparticular in form of their alkali salts, as well as protein fatty acidcondensates.

Examples of suitable nonionic surfactants include, but are not limitedto, fatty alcohol acid or amide ethoxylates, monoglyceride ethoxylates,sorbitan ester ethoxylates alkyl polyglycosides, mixtures thereof, andthe like. Certain preferred nonionic surfactants include polyoxyethylenederivatives of polyol esters, wherein the polyoxyethylene derivative ofpolyol ester (1) is derived from (a) a fatty acid containing from about8 to about 22, and preferably from about 10 to about 14 carbon atoms,and (b) a polyol selected from sorbitol, sorbitan, glucose, α-methylglucoside, polyglucose having an average of about 1 to about 3 glucoseresidues per molecule, glycerine, pentaerythritol and mixtures thereof,(2) contains an average of from about 10 to about 120, and preferablyabout 20 to about 80 oxyethylene units; and (3) has an average of about1 to about 3 fatty acid residues per mole of polyoxyethylene derivativeof polyol ester. Examples of such preferred polyoxyethylene derivativesof polyol esters include, but are not limited to PEG-80, sorbitanlaurate, and Polysorbate 80.

Another class of suitable nonionic surfactants includes long chain alkylglucosides or polyglucosides, which are the condensation products of (a)a long chain alcohol containing from about 6 to about 22, and preferablyfrom about 8 to about 14 carbon atoms, with (b) glucose or aglucose-containing polymer. Preferred alkyl gluocosides comprise fromabout 1 to about 6 glucose residues per molecule of alkyl glucoside. Apreferred glucoside is decyl glucoside, which is the condensationproduct of decyl alcohol with a glucose polymer and is availablecommercially from Cognis Corporation of Ambler, Pa. under the tradename,“Plantaren 2000.”

Various cationic surfactants may also be suitable for use in the presentcompositions. Examples of suitable cationic surfactants include, but arenot limited to alkyl quaternaries (mono, di, or tri), benzylquaternaries, ester quaternaries, ethoxylated quaternaries, alkylamines, and mixtures thereof, wherein the alkyl group has from about 6carbon atoms to about 30 carbon atoms, with about 8 to about 22 carbonatoms being preferred.

Also possible is the incorporation of amphoteric surfactants, such asalkyl betaines, alkyl amido betaines, and alkyl amphoacetates.

To still further improve the stability, the liquid pharmaceuticalcompositions may also include one or more anti-oxidants. For example,hydrophobic anti-oxidants include butylated hydroxytoluene, butylatedhydroxyanisole, propyl gallate, and α.-tocopherol, DL-tocopherol,α-tocopherol acetate, α-tocopherol Tocopherol Polyethylene GlycolSuccinate (Vitamin E TPGS), L-cysteine, or hydrophilic anti-oxidants,including sodium EDTA and thioglycerol. Most typically, theconcentration of the anti-oxidant will be between 0.005% and 5%weight/weight of the total composition.

Additionally, or alternatively, the contemplated liquid pharmaceuticalcompositions may include a preservative (e.g., phenol, thimerosal,chlorobutanol, benzyl alcohol, m-cresol, phenoxyethanol, methylparabenand propylparaben), typically at a concentration of between 0.001%weight/weight and about 5% weight/weight of the total composition, andis most typically between about 0.003% and about 2.0% weight/weight ofthe total composition.

The liquid pharmaceutical compositions contemplated herein may furtherinclude isotonicity agents and/or lyoprotectants. Suitable isotonicityagents include sodium chloride, glycerol, and thioglycerol. Suitablelyoprotectants are described above.

The liquid pharmaceutical compositions can be prepared according to amethod comprising the steps:

(a) placing a desired amount of a peptide epoxy ketone in a compoundingvessel;

(b) adding one or more water miscible pharmaceutically acceptableorganic solvents suitable for injection to the compounding vessel;

(c) optionally adding water to the compounding vessel;

(d) mixing the contents of the compounding vessel;

(e) optionally adding one or more excipients to the compounding vessel;

(f) adding a non-volatile sugar acid to the compounding vessel; and

(g) mixing the contents of the compounding vessel until the non-volatilesugar acid is dissolved.

The contents of the compounding vessel may be sterilized using any knownmethods of sterilization, including filtration through 0.22 micronfilters (e.g., PVDF filters), heat sterilization, radiation (e.g.,gamma, electron beam, microwave), and/or ethylene oxide sterilization.

The pharmaceutical compositions of the instant invention are suitablefor administration to mammals, including humans. When administered tohumans, the compositions are typically administered by intramuscular,subcutaneous, intra-arterial, or oral routes.

Regardless of the particular composition, it is preferred that thecomposition is packaged in a container suitable for single or multi-use.Such containers include an ampoule, a vial, a pre-filled syringe, and anintravenous bag. Multi-use containers may contain the peptide epoxyketone in an amount suitable to allow at least two distinct uses, moretypically at least five distinct uses, and even more typically at leastten distinct uses (each use which may or may not require the samequantity of composition administered to the patient). Thus, preferredmulti-use containers will be configured to contain a volume of thecomposition that is suitable for multiple and independentadministrations. Such containers include vials with a rubber stopperthat can be pierced with a needle of a syringe.

The following examples are presented for illustrative purposes only, andare not intended to limit the scope of the invention.

EXAMPLES

For each of the Examples described below, the following HPLC methodologywas used to determine the relative retention time (RRT) of impurities inthe compositions:

Mobile Phase A: 0.5 mL of TFA in 1000 mL of distilled water

Mobile Phase B: 100% HPLC grade Acetonitrile

Diluent: Acetonitrile: Water (1:1 v/v)

Elution program:

Time Min Flow mL % A % B 0.01 1.00 95.0 5.0 5.00 1.00 90.0 10.0 10.001.00 85.0 15.0 38.00 1.00 20.0 80.0 40.00 1.00 0.0 100.0 43.00 1.00 0.0100.0 45.00 1.00 95.0 5.0 50.00 1.00 95.0 5.0

Flow rate: 1.0 mL/min

Detector Wavelength: 210 nm

Injection volume: 10 micro liters

Run time: 50 min

Column: Waters Symmetry C18 (250×4.6 mm), 5 microns

Sample temperature: 5° C.

Column temperature: 25° C.

Example 1 Ready-to-Dilute Composition (60 mg/Vial) to be Diluted BeforeInjecting

Ingredient Amount Carfilzomib  60 milligrams Ethanol  5 millilitersPropylene Glycol  2.5 milliliters Lactobionic Acid 400 milligramsAcetate Buffer (15 millimolar, pH 3.5)  1 milliliter Polysorbate 80 500milligrams 5% Dextrose  0.5 milliliters

The composition of Example 1 was prepared by placing 60 milligrams ofCarfilzomib in a compounding vessel. 5 milliliters of ethanol was thenadded to the compounding vessel and the contents of the compoundingvessel were mixed. 2.5 milliliters propylene glycol was added to thecompounding vessel and the contents of the vessel were again mixed.Next, 1 milliliter of 15 mM acetate buffer and 500 milligrams ofPolysorbate 80 were added to the compounding vessel and the contentstherein were mixed. 400 milligrams lactobionic acid was then added tothe compounding vessel and the contents of the vessel were mixed untilthe lactobionic acid was dissolved. 0.5 milliliters of a 5% dextrosesolution was then added to the compounding vessel and the contentstherein were mixed. Once mixing was completed, the contents of thecompounding vessel were filtered through a 0.22 micron PVDF filter.

Physicochemical data for the composition of Example 1 is shown below:

Impurities Assay RRT RRT RRT RRT Temperature Time Appearance (%) 0.820.98 1.02 1.18 Total T = 0 CCS 104.9 — 0.79 0.77 — 1.56 40° C./75% RH 1Week CCS 89.3 0.05 1.26 0.77 — 2.05 2 Week CCS 113.4 0.57 2.11 0.85 —3.53 1 Month CCS 90.6 0.10 2.67 0.64 4.23 7.64 3 Month CCS 44.64 0.284.07 0.28 16.15 20.78 25° C./60%RH 1 Month CCS 105.8 0.15 1.11 0.77 —2.03 3 Month CCS 107.9 0.63 2.13 0.73 1.22 4.71 CCS = clear colorlesssolution RRT = relative retention time

As can be seen from the physicochemical data, the composition of Example1, which comprises lactobionic acid as well as ethanol and propyleneglycol solvents, remained stable for an extended period of time evenwhen stored at temperatures of 25° Celsius. The ready to dilutecomposition is stable at room temperature for at least 3 months.

Example 2 Ready-to-Dilute Composition (60 mg/Vial) to be Diluted BeforeInjecting

Ingredient Amount Carfilzomib  60 milligrams Ethanol  5 milliliters PEG300  2.5 milliliters Lactobionic Acid 400 milligrams Acetate Buffer (15millimolar, pH 3.5)  1 milliliter Polysorbate 80 500 milligrams 5%Dextrose  0.5 milliliters

The composition of Example 2 was prepared by placing 60 milligrams ofCarfilzomib in a compounding vessel. 5 milliliters of ethanol was thenadded to the compounding vessel and the contents of the compoundingvessel were mixed. 2.5 milliliters PEG 300 was added to the compoundingvessel and the contents of the vessel were again mixed. Next, 1milliliter of 15 mM acetate buffer, and 500 milligrams of Polysorbate80, were added to the compounding vessel and the contents therein weremixed. 400 milligrams lactobionic acid was then added to the compoundingvessel and the contents of the vessel were mixed until the lactiobionicacid was dissolved. 0.5 milliliters of a 5% dextrose solution was thenadded to the compounding vessel and the contents therein were mixed.Once mixing was completed, the contents of the compounding vessel werefiltered through a 0.22 micron PVDF filter.

Physicochemical data of the composition of Example 2 is shown below:

Impurities Assay RRT RRT RRT RRT Temperature Time Appearance (%) 0.820.98 1.02 1.18 Total T = 0 CCS 97.0 — 0.78 0.77 — 1.55 40° C./75% RH 1Week CCS 98.1 0.34 1.35 0.77 — 2.45 2 Week CCS 87.0 0.66 1.96 0.71 —3.33 1 Month CCS 84.8 1.33 2.83 0.57 2.93 7.66 3 Month CCS 53.0 0.224.19 0.37 8.74 13.52 25° C./60% RH 1 Month CCS 106.0 0.22 1.14 0.75 —2.08 3 Month CCS 99.65 0.84 2.14 0.65 0.66 4.29 CCS = clear colorlesssolution PPT = precipitate RRT = relative retention time

As can be seen from the physicochemical data, the composition of Example2, which comprises lactobionic acid as well the solvents ethanol and PEG300, remained stable for an extended period of time even when stored attemperatures of 25° Celsius. The ready to dilute composition is stableat room temperature for at least 3 months.

Example 3 Ready-to-Use Composition (2 mg/mL)

Ingredient Amount Carfilzomib 60 mg Ethanol 12 mL PEG 300 2.5 mLLactobionic Acid 400 mg Acetate Buffer (15 millimolar, pH 3.5) 2 mLPolysorbate 80 500 mg 5% Dextrose Q.S to 30 g

The composition of Example 3 was prepared by placing 60 milligrams ofCarfilzomib in a compounding vessel. 12 milliliters of ethanol was thenadded to the compounding vessel and the contents of the compoundingvessel were mixed. 2.5 milliliters PEG 3000 was added to the compoundingvessel and the contents of the vessel were again mixed. Next, 2 mL of 15mM acetate buffer and 500 milligrams of Polysorbate 80 were added to thecompounding vessel and the contents therein were mixed. 400 milligramslactobionic acid was then added to the compounding vessel and thecontents of the vessel were mixed until the lactobionic acid wasdissolved. A sufficient amount of 5% dextrose solution was then added tothe compounding vessel so as to provide 30 g and the contents thereinwere mixed. Once mixing was completed, the contents of the compoundingvessel were filtered through a 0.22 micron PVDF filter.

Physicochemical data for the composition of Example 3 is shown below:

Impurities Assay RRT RRT RRT RRT Temperature Time Appearance (%) 0.820.98 1.02 1.18 Total T = 0 CCS 107.5 — 0.85 0.77 — 1.61 40° C./75% RH 1Week CCS 100.9 0.47 1.7 0.77 — 2.91 2 Week CCS 96.2 0.82 2.47 0.75 4.031 Month CCS 88.4 1.79 3.75 0.6 — 6.15 2 Month CCS 105.5 0.39 1.21 0.64 —2.23 25° C./60% RH 1 Month CCS 108.1 0.48 1.32 0.75 — 2.55 2 Month CCS99.31 0.96 2.53 0.56 — 4.04 3 Month CCS 97.46 1.07 2.70 0.55 — 4.31 4°C. 2 Month CCS 113.17 0.41 1.33 0.69 — 2.43 3 Month CCS 108.29 — 0.790.66 — 1.44 CCS = clear colorless solution RRT = relative retention time

The ready to use composition is stable at room temperature for at least3 months.

The preceding examples, and exemplary language, are merely illustrateand should not be construed to limit the scope of the inventiondiscussed herein, unless otherwise claimed. In addition, no language inthe specification should be construed as indicating any non-claimedelement as essential to the practice of the invention. For instance,while particular embodiments of the present invention have beenillustrated and described, it should be apparent to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. For instance, allmethods described herein can be performed in any suitable order unlessotherwise indicated herein or otherwise clearly contradicted by context.

1. A pharmaceutical composition, comprising: (i) a peptide epoxy ketoneor a pharmaceutically acceptable salt thereof; (ii) a solvent systemcomprising a water miscible pharmaceutically acceptable organic solventsuitable for injection selected from the group consisting of ethanol,propylene glycol, polyethylene glycol, and mixtures thereof, andoptionally water; and (iii) a non-volatile sugar acid.
 2. Thepharmaceutical composition of claim 1, wherein the non-volatile sugaracid is selected from the group consisting of lactobionic acid, glycolicacid, malic acid, citric acid, lactic acid, mandelic acid, and tartaricacid.
 3. The pharmaceutical composition of claim 2, wherein the peptideepoxy ketone is carfilzomib or a pharmaceutically acceptable saltthereof.
 4. The pharmaceutical composition of claim 1, wherein thesolvent system comprises ethanol and propylene glycol.
 5. Thepharmaceutical composition of claim 1, wherein the solvent systemcomprises ethanol and polyethylene glycol.
 6. The pharmaceuticalcomposition of claim 1, wherein peptide epoxy ketone is carfilzomib or apharmaceutically acceptable salt thereof and the non-volatile sugar acidis lactobionic acid.
 7. The pharmaceutical composition of claim 6,wherein the concentration of the peptide epoxy ketone in the solventsystem ranges from about 0.1 to about 10.0 mg/mL and the weight ratio ofthe peptide epoxy ketone to lactobionic acid ranges from about 2:1 to15:1.
 8. The pharmaceutical composition of claim 7, wherein the solventsystem comprises ethanol, propylene glycol, and water.
 9. Thepharmaceutical composition of claim 7, wherein the solvent systemcomprises ethanol, polyethylene glycol, and water.
 10. Thepharmaceutical composition of claim 1 comprising: (i) carfilzomib or thepharmaceutically acceptable salt thereof; (ii) lactobionic acid, whereinthe weight ratio of the lactobionic acid to carfilzomib ranges from 2:1to 15:1; and (iii) a solvent system comprising ethanol, propyleneglycol, and water, wherein the ethanol and propylene glycol comprisemore than about 50% (v/v) of the solvent system, wherein theconcentration of carfilzomib ranges from about 1 to about 6 mg/mL. 11.The pharmaceutical composition of claim 1 comprising: (i) carfilzomib orthe pharmaceutically acceptable salt thereof; (ii) lactobioic acid,wherein the weight ratio of the lactobionic acid to carfilzomib rangesfrom about 4:1 to 7:1; and (iii) a solvent system comprising ethanol,polyethylene glycol, and water, wherein the ethanol and polyethyleneglycol comprise more than about 50% (v/v) of the solvent system, whereinthe concentration of the carfilzomib ranges from about 1 to about 6mg/mL.
 12. A pharmaceutical composition comprising a peptide epoxyketone and a sugar, wherein the pharmaceutical composition islyophilized.
 13. The pharmaceutical composition of claim 12, wherein thesugar acid is lactobionic acid.
 14. The pharmaceutical composition ofclaim 12, wherein the peptide epoxy ketone is carfilzomib.
 15. Thepharmaceutical composition of claim 14, sugar acid is lactobionic acid.16. The pharmaceutical composition of claim 13, wherein the weight ratioof the peptide epoxy ketone to lactobionic acid ranges from about 2:1 to10:1.
 17. The pharmaceutical composition of claim 15, wherein the weightratio of the lactobionic acid to carfilzomib ranges from about 2:1 to10:1.